Topical doxyl composition and method

ABSTRACT

A composition and method for ameliorating a cellular dysfunction of a tissue such as the cosmetic treatment of hair loss and stimulation of hair growth is disclosed. The method comprises administering 2,2-disubstituted-4,4-dimethyl-3-oxazolidinyloxy (DOXYL) to the affected tissue.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application is a continuation-in-part of application Ser. Nos.08/229,374 filed Apr. 18, 1994, now U.S. Pat. No. 5,470,876, and08/193,228, filed Feb. 7, 1994 now U.S. Pat. No. 5,472,687, which arecontinuations-in-part of application Ser. No. 08/021,970, filed Feb. 24,1993, now U.S. Pat. No. 5,352,442; which is a continuation-in-part ofapplication Ser. No. 07/149,720, filed Jan. 29, 1988, abandoned; whichis a continuation-in-part of application Ser. No. 07/008,186, filed Jan.28, 1987, abandoned; which is a continuation-in-part of application Ser.No. 06/858,050, Apr. 30, 1986, abandoned; which is acontinuation-in-part of application Ser. No. 06/757,131, Jul. 18, 1985,abandoned.

FIELD OF THE INVENTION

This invention relates to a topical composition comprising4,4-dimethyl-3-oxazolidinyloxy (DOXYL) and a method for treating hairloss therewith.

BACKGROUND OF THE INVENTION

Several compounds have recently gained recognition for amelioratingcellular dysfunction. One type of dysfunction which has been wellstudied is alopecia for which anti-alopecia agents such as minoxidil andcyoctol have gained attention. However, most of these anti-alopeciaagents are only minimally effective in some cases and/or can causeadverse dermatological or systemic reactions. Thus, the search continuesfor new, safer and more effective anti-alopecia agents as well as agentsuseful for treating other dysfunctionalities.

SUMMARY OF THE INVENTION

Applicant has discovered that2,2-disubstituted-4,4-dimethyl-3-oxazolidinyloxy (DOXYL) has propertiesin the body for ameliorating cellular dysfunction in tissue attributed,in part, to high energy oxygen and hydroxyl free radicals, and enhancingrecuperation of the tissue.2,2-Disubstituted-4,4-dimethyl-3-oxazolidinyloxy can be administered,for example, as an anti-alopecia agent to stimulate cosmetic hairgrowth, or as a protectant to minimize hair loss during cancertreatments known to induce hair loss.

DETAILED DESCRIPTION OF THE INVENTION

In the present invention,2,2-disubstituted-4,4-dimethyl-3-oxazolidinyloxy (DOXYL) is compoundedin a pharmaceutical formulation or carrier for topical or internaladministration. The topical pharmaceutical carrier in which the DOXYL isgenerally substantially homogeneously dispersed can be an aqueousdispersion or suspension, or a water-in-oil or oil-in-water emulsiondepending on the administration route. Topical pharmaceutical carrierswhich can be mentioned include water, urea, alcohols and glycols such asmethanol, ethanol, propanol, butanol, ethylene glycol, propylene glycol,and the like. Internally administered pharmaceutical carriers typicallyinclude a sterile vehicle such as water or ethanol in which the DOXYL issuspended, dispersed or dissolved.

Suitable water-in-oil emulsions are commercially available under thedesignations Aquaphor, cold cream, Eucerin, hydrous lanolin, Hydrosorbhydrophilic petrolatum, Nivea, Polysorb, Qualatum and Velvachol.Suitable oil-in-water emulsions are available commercially under thedesignations acid mantle cream, Almay emulsion cream, Cetaphil,Dermabase, Dermavan, hydrophilic ointment, Keri cream, Lubriderm cream,Multibase cream, Neobase cream, Unibase cream, Vanibase cream and Wibi.The carrier may further contain various other emollients, emulsifiers,water, perfumes, colorants, preservatives, and the like. The topicalformulation is in the form of a cream, lotion, shampoo, cream rinse, orthe like.

DOXYL is a stable nitroxide radical which is a commercially availablespin label. DOXYL is typically formed by the condensation of2-amino-2-methyl-1-propanol and a ketone, followed by oxidation. Thecarbon in the carbonyl moiety of the ketone becomes the 2-positioncarbon in the DOXYL. Thus, the DOXYL group is typically disubstituted inthe 2-position. For example, 12-doxylstearic acid is formed fromcondensation and subsequent oxidation of 12-ketostearic acid and2-amino-2-methyl-1-propanol. Another name for 12-doxylstearic acid wouldbe 2-hexyl-2- 11-hydroxy-11-oxoundecyl!-4,4-dimethyl-3-oxazolidinyloxy.Similarly, using methylethyl ketone would form2-ethyl-2,4,4-trimethyl-3-oxazolidinyloxy, which is also referred to inthe art as OXANO. Exemplary 2,2-disubstituted doxyl radicals include3-doxyl-5α-cholestane, 3-doxyl-17β-hydroxy-5α-androstane, 5-doxylstearicacid, 7-doxylstearic acid, 12-doxylstearic acid, 16-doxylstearic acid,5-doxylstearic acid methyl ester, 7-doxylstearic acid methyl ester,12-doxylstearic acid methyl ester, 16-doxylstearic acid methyl ester,and the like.

Effective amounts of the DOXYL generally range from about 0.01 to about20% by weight of the administered composition, more preferably fromabout 0.1 to about 10% by weight, most preferably from about 0.5 toabout 3% by weight, but more or less can be present in the compositiondepending on the particular DOXYL formulation and the treatmentconditions.

The DOXYL can be used alone or in combination with other additamentswhich are available to enhance the function of hair growth stimulationsuch as, for example, the hydroxyl radical scavengers, antiandrogens andothers described in International Publication No. WO 87/00427(International Application No. PCT/U.S. application Ser. No. 86/01393)published on Jan. 29, 1987; and European Patent Application No.89300785.6, Publication No. 0327263/A1, published Aug. 9, 1989; both ofwhich are hereby incorporated in their entirety herein as though fullyset forth verbatim, including reference therein to the publication ofRoss & Ross, "Selected Specific Rates of Reactions of Transients FromWater In Aqueous Solution. III. Hydroxyl Radical and Pure HydroxylRadicals and Their Radical Ions," National Standard Reference DataSeries, National Bureau of Standards, 59 (1977), which is alsoincorporated herein by reference.

According to the present invention, the DOXYL can be administered to theskin to be treated, such as the scalp. Depending on the type of hairloss or alopecia being treated and the conditions thereof, thestimulation of hair growth can usually be obtained by topicalapplication, preferably repeated daily application for a period of 3-6months. The utility of topically applied DOXYL is not limited thereto,however, and the stimulation of hair growth can include an increasedrate of growth, increased hair diameter, follicular neogenesis, and thelike; inhibiting hair loss or alopecia from progressing, for example, inmale pattern baldness, or during the course of treatment with othertherapeutic agents known to induce hair loss, such as chemotherapy orradiation therapy in cancer treatment. DOXYL can also be usefuladministered topically, orally or parentally in ameliorating the rate ofprotein oxidation, DNA scission, cell viability loss, and the like inthe tissue of internal organs such as the heart and brain; andameliorating capillary loss, tissue atrophy characterized by a decreasein collagen and/or elastin and a decreased number, size and reproductionpotential of fibroblasts; and strengthening the dermal-epidermaljunction in skin; ischemic reperfusion injury secondary to myocardialinfarction, stroke and surgical procedures; wound healing, for example,in burns and diabetic ulcerations; inflammatory and degenerativediseases such as rheumatoid arthritis, lupus and the like; fibroticdiseases such as Peyronie's disease, scarring, pulmonary fibrosis, andvitreous fibrosis; prevention of free-radical-induced vascular damagesuch as in atherosclerosis; other free radical diseases as outlined inProctor et al., "Free Radicals and Disease in Man," PhysiologicalChemistry and Physics and Medical NMR, volume 16, pp. 175-195 (1984)which is hereby incorporated herein by reference; and the like.

The invention is illustrated by way of the following examples:

EXAMPLE 1

A DOXYL shampoo is prepared by mixing 0.5 g of 5-doxylstearic acid in500 ml of a commercially available shampoo. The shampoo is used daily onthe scalp for normal shampooing of the hair for a period of from 3 to 6months to obtain cosmetic hair growth.

EXAMPLE 2

A solution of 5-doxylstearic acid is prepared and used in the course ofradiation treatment. 5-Doxylstearic acid, obtained commercially fromAldrich Chemical Company, is dissolved in 70 percent ethanol/30 percentwater at a concentration of 70 mg/ml. Topical application of thesolution is made prior to irradiation exposure at 20Gy to 50Gy. Hairloss in the treated DOXYL subjects is less severe and returns to normalmore rapidly than in the control group similarly treated with the sameethanol/water solution without DOXYL. Skin samples obtained from thetreated group test positive for the presence of 5-doxylstearic acid,while other tissue and blood specimens generally test negative. Theapplication of the solution can also continue daily after theirradiation exposure. See Goffman, et al., "Topical Application ofNitroxide Protects Radiation-Induced Alopecia in Guinea Pigs,"International Journal of Radiation Oncology, Biology and Physics, Volume22, pp. 803-806, 1992, which is hereby incorporated herein by reference.

EXAMPLE 3

A 0.4 or 1 mM solution of DOXYL is used to significantly reduce cardiacinjury caused by reperfusion arrhythmia-ventricular fibrillation andventricular tachycardia, as well as, post ischemic release of lactatedehydrogenase and OH-- formation in isolated rat hearts subjected toregional ischemia. The rat hearts are obtained and perfused using amodified Krebs-Henseleit (KH) buffer, as detailed in Gelvan et al.,"Cardiac Reperfusion Damage Prevented by a Nitroxide Free Radical,"Proceedings of the National Academy of Sciences, USA, Medical Sciences,Vol. 88, pp. 4680-4684, June 1991, which is hereby incorporated hereinby reference, in which a TEMPO solution was added to the perfusate.After reperfusion, heart function and resulting damage is analyzed.DOXYL is found to strongly protect against reperfusion injury bypreventing OH-- formation rather than by decreasing heart rate or bydirect suppression of arrhythmia.

EXAMPLE 4

The protective effect of OXANO against oxidative damage is shown byexposing to H₂ O₂ the E. coli xthA DNA repair-deficient mutant which ishypersensitive to H₂ O₂, and assaying oxidative damage by monitoringcell survival, as described in Samuni et al., "Nitroxides block DNAscission and protect cells from oxidative damage," Biochemistry, vol.30, pp. 555-561 (1991) which is hereby incorporated herein by reference.The presence of OXANO in the incubation medium protects both growing andresting cells from H₂ O₂ toxicity without lowering the H₂ O₂concentration.

EXAMPLE 5

The protective effect of spirocyclohexyldoxyl (CHD) and TEMPOL againstoxidative damage is shown by incubating Chinese hamster V79 cellsexposed to H₂ O₂ in the presence of CHD or TEMPOL as described in WO91/13619 published Sept. 19, 1991, which is hereby incorporated hereinby reference. The presence of 5 mM CHD or TEMPOL fully protects thecells against H₂ O₂ cytotoxicity.

The invention is described above and illustrated herein with referenceto specific chemical formulas, preparations and therapeutic and cosmeticapplications. Many variations and modifications will become apparent tothose skilled in the art in view of the foregoing disclosure. It isintended that the following claims are not to be limited thereby, andare to be construed in accordance with the spirit and scope thereof.

I claim:
 1. A method for inhibiting the activity of oxygen and hydroxylfree radicals in tissue of an organism, comprising the stepof:administering 2,2-disubstituted-4,4-dimethyl-3-oxazolidinyloxy to thetissue in an amount effective to inhibit the free radicals.
 2. Themethod of claim 1, wherein the administration step is topical.
 3. Themethod of claim 2, wherein the2,2-disubstituted-4,4-dimethyl-3-oxazolidinyloxy is in the form of adispersion, suspension or emulsion selected from creams, lotions,shampoos and cream rinses.
 4. The method of claim 3, wherein thedispersion, suspension or emulsion comprises from about 0.01 to about 20percent by weight of said oxazolidinyloxy.
 5. The method of claim 1,wherein the 2,2-disubstituted-4,4-dimethyl-3-oxazolidinyloxy is selectedfrom 3-doxyl-5α-cholestane, 3-doxyl-17β-hydroxy-5α-androstane,5-doxylstearic acid, 7-doxylstearic acid, 12-doxylstearic acid,16-doxylstearic acid, 5-doxylstearic acid methyl ester, 7-doxylstearicacid methyl ester, 12-doxylstearic acid methyl ester, and16-doxylstearic acid methyl ester.
 6. A method for treating hair losscomprising the step of:administering an effective amount of substitutedor unsubstituted 2,2disubstituted-4,4-dimethyl-3-oxazolidinyloxy to theaffected area.
 7. The method of claim 6, wherein the administration stepis topical.
 8. The method of claim 7, wherein the2,2-disubstituted-4,4-dimethyl-3-oxazolidinyloxy is in the form of adispersion, suspension or emulsion selected from creams, lotions,shampoos and cream rinses.
 9. The method of claim 6, whereinthe2,2-disubstituted-4,4-dimethyl-3-oxazolidinyloxy is selected from3-doxyl-5α-cholestane, 3-doxyl-17β-hydroxy-5α-androstane, 5-doxylstearicacid, 7-doxylstearic acid, 12-doxylstearic acid, 16-doxylstearic acid,5-doxylstearic acid methyl ester, 7-doxylstearic acid methyl ester,12-doxylstearic acid methyl ester, and 16-doxylstearic acid methylester.
 10. The method of claim 9, wherein the dispersion, suspension oremulsion comprises from about 0.01 to about 20 percent by weight of saidoxazolidinyloxy.
 11. A topical pharmaceutical composition useful for thecosmetic treatment of hair loss, comprising a2,2-disubstituted-4,4-dimethyl-3-oxazolidinyloxy in association with atopical pharmaceutical carrier comprising a water and oil emulsion. 12.The topical pharmaceutical composition of claim 11, comprising from 0.01to 20 weight percent of an oxazolidinyloxy selected from3-doxyl-5α-cholestane, 3-doxyl-17β-hydroxy-5α-androstane, 5-doxylstearicacid, 7-doxylstearic acid, 12-doxylstearic acid, 16-doxylstearic acid,5-doxylstearic acid methyl ester, 7-doxylstearic acid methyl ester,12-doxylstearic acid methyl ester, and 16-doxylstearic acid methylester.
 13. A topical pharmaceutical composition useful for the cosmetictreatment of hair loss comprising a2,2-disubstituted-4,4-dimethyl-3-oxazolidinyloxy in association with atopical pharmaceutical carrier selected from creams, lotions, shampoosand cream rinses.
 14. The topical pharmaceutical composition of claim13, comprising from 0.01 to 20 weight percent of an oxazolidinyloxyselected from 3-doxyl-5α-cholestane, 3-doxyl- 17β-hydroxy-5α-androstane,5-doxylstearic acid, 7-doxylstearic acid, 12-doxylstearic acid,16-doxylstearic acid, 5-doxylstearic acid methyl ester, 7-doxylstearicacid methyl ester, 12-doxylstearic acid methyl ester, and16-doxylstearic acid methyl ester.